NDLI: Large-scale zebrafish embryonic heart dissection for transcriptional analysis.

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Large-scale zebrafish embryonic heart dissection for transcriptional analysis.

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Large-scale zebrafish embryonic heart dissection for transcriptional analysis.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lombardo, Verónica A. Otten, Cécile Abdelilah-Seyfried, Salim
Description	Author Affiliation: Lombardo VA ( Max Delbrück Center for Molecular Medicine); Otten C ( Max Delbrück Center for Molecular Medicine); Abdelilah-Seyfried S ( Max Delbrück Center for Molecular Medicine)
Abstract	The zebrafish embryonic heart is composed of only a few hundred cells, representing only a small fraction of the entire embryo. Therefore, to prevent the cardiac transcriptome from being masked by the global embryonic transcriptome, it is necessary to collect sufficient numbers of hearts for further analyses. Furthermore, as zebrafish cardiac development proceeds rapidly, heart collection and RNA extraction methods need to be quick in order to ensure homogeneity of the samples. Here, we present a rapid manual dissection protocol for collecting functional/beating hearts from zebrafish embryos. This is an essential prerequisite for subsequent cardiac-specific RNA extraction to determine cardiac-specific gene expression levels by transcriptome analyses, such as quantitative real-time polymerase chain reaction (RT-qPCR). The method is based on differential adhesive properties of the zebrafish embryonic heart compared with other tissues; this allows for the rapid physical separation of cardiac from extracardiac tissue by a combination of fluidic shear force disruption, stepwise filtration and manual collection of transgenic fluorescently labeled hearts.
File Format	HTM / HTML
e-ISSN	1940087X
Journal	Journal of Visualized Experiments
Issue Number	95
Language	English
Publisher	MyJove Corp.
Publisher Date	2015-01-12
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Cardiac Surgical Procedures Veterinary Gene Expression Profiling Heart Embryology Myocardium Chemistry Rna, Messenger Isolation & Purification Zebrafish Surgery Animals Animals, Genetically Modified Dissection Real-time Polymerase Chain Reaction Zebrafish Proteins Genetics Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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